食欲素在细胞氧化应激中的神经保护作用及其机制研究  

Research on the neuroprotective roles of Orexin in cell model of oxidative stress

作　　者：李迎帅 曹飞 王春梅[1] 杨春青 刘霞 王琴琴 LI Yingshuai;CAO Fei;WANG Chunmei;YANG Chunqing;LIU Xia;WANG Qinqin(School of Mental Health,Jining 272000,China;College of Rehabilitation Medicine,Jining Medical University,Jining 272000,China)

机构地区：[1]济宁医学院精神卫生学院,济宁272000 [2]济宁医学院康复医学院,济宁272000

出　　处：《济宁医学院学报》2022年第5期335-338,343,共5页Journal of Jining Medical University

基　　金：2020年山东省大学生创新创业训练计划项目(S202010443058);2020年济宁医学院大学生创新训练计划项目(cx2020025);山东省自然科学基金项目(ZR2021QC039);山东省医药卫生科技发展计划项目(202002 081082)。

摘　　要：目的 探讨食欲素(Orexin)在氧化应激中的具体作用及其机制。方法 培养海马神经元细胞系HT-22细胞,随机分为对照组、H_(2)O_(2)(hydrogen peroxide)组、OA(Orexin A)组、OB(Orexin B)组、H_(2)O_(2)+OA组及H_(2)O_(2)+OB组,通过H_(2)O_(2)处理HT-22细胞建立氧化应激体外模型,采用显微镜分析方法检测各组细胞数量变化;利用实时荧光定量PCR(Quantitative Real-time PCR,qPCR)及蛋白印迹(Westernblotting)等方法探究转录因子Nrf2在HT-22细胞氧化应激模型中的表达水平及其作用。结果 与对照组相比,H_(2)O_(2)显著增强HT-22细胞的死亡(P<0.05);给予OA及OB处理后,与H_(2)O_(2)组相比,OA及OB均显著降低H_(2)O_(2)引起的HT-22细胞的死亡(P<0.05);与对照组相比,H_(2)O_(2)处理的HT-22细胞Nrf2的基因表达水平显著升高(P<0.05);与H_(2)O_(2)组相比,OA和OB显著增加HT-22细胞内的Nrf2的表达水平(P<0.05)。结论 食欲素可通过增强HT-22细胞内Nrf2的表达水平降低H_(2)O_(2)引起的HT-22细胞的神经损伤,进而发挥神经保护作用。Objective To explore the roles and the underlying mechanisms of orexin in oxidative stress.Methods HT-22 cells were randomly divided into control group,H_(2)O_(2)(hydrogen peroxide) group,OA(Orexin A) group,OB(Orexin B)group,H_(2)O_(2)+OA group and H_(2)O_(2)+OB group.The cell model of oxidative stress was constructed by treating HT-22 cells with H_(2)O_(2).OA and OB were added to investigate the effects of orexin on cell number and morphology using microscopic analysis.The levels of Nrf2 in HT-22 cells were detected by qPCR(Quantitative Real-time PCR) and Western blotting.Results Compared with the control group,the cell number decreased significantly after H_(2)O_(2) treatment(P<0.01);OA and OB couldrobustly decrease the death of HT-22 cells induced by H_(2)O_(2)(P<0.05);Compared with the control group,H_(2)O_(2) led to a significant increase of the expression level of Nrf2 in HT-22 cells(P<0.05);Furthermore,compared with the H_(2)O_(2) group,OA and OB obviously enhanced the expression levels of Nrf2 in HT-22 cells.Conclusion All of these results suggested that orexin in creased the levels of Nrf2 in HT-22 cells and decreased the injury of HT-22 cells induced by H_(2)O_(2),leading to the neuroprotective roles in oxidative stress.

关 键 词：食欲素 氧化应激 HT-22细胞 过氧化氢 NRF2 

分 类 号：R966[医药卫生—药理学]